| T7 RNA聚合酶表达系统的真核化及其偶联表达系统的建立 |
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| 引用本文： | 郑海学,靳野,尹双辉,郭慧琛,尚佑军,白兴文,刘湘涛,谢庆阁.T7 RNA聚合酶表达系统的真核化及其偶联表达系统的建立[J].生物工程学报,2007,23(5):947-952. |
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| 作者姓名： | 郑海学 靳野 尹双辉 郭慧琛 尚佑军 白兴文 刘湘涛 谢庆阁 |
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| 作者单位： | 1. 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,兰州,730046;中国农业科学院研究生院,北京,100081
2. 中国农业科学院兰州兽医研究所家畜疫病病原生物学国家重点实验室,农业部畜禽病毒学重点开放实验室,兰州,730046 |
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| 基金项目： | 国家重点基础研究发展计划(973计划);国家科技攻关计划 |
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| 摘 要： | 为了使T7RNA聚合酶(T7RNAP)表达系统真核化,首先,利用两种方法建立能够表达T7RNAP的真核细胞:(1)共转染表达T7RNAP的真核表达重组质粒于靶细胞;(2)利用稳定表达T7RNAP的BHK-21细胞系。然后,将FMDV内部核糖体进入位点(IRES)序列和增强型绿色荧光蛋白(EGFP)基因定向克隆进原核载体pET-40a-c( )的T7启动子下游,得到的重组质粒pIERS-EGFP-E。用该质粒分别转染上述两种细胞,在紫外显微镜下都能够观察到绿色荧光,表明真核化的T7RNAP偶联表达系统建立。并利用流式细胞仪对偶联表达水平进行了分析。这为利用该系统真核高效表达外源蛋白奠定了坚实的基础。用实验证明了FMDV5′端含有IERS具有介导非帽依赖性的翻译的功能,为以IERS为基础的双顺反子表达系统的建立及深入研究IERS与相关蛋白的功能奠定了基础。T7RNAP偶联表达体系是一种良好的外源基因表达体系。
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| 关 键 词： | 口蹄疫病毒 T7 RNA聚合酶表达系统 偶联表达系统 |
| 文章编号： | 1000-3061（2007）05-0947-06 |
| 修稿时间： | 2007-01-16 |
Eukaryonization of T7 RNA Polymerase Prokaryotic Expression System and Development of Its Couple Expression System |
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| ZHENG Hai-Xue,JIN Ye,YIN Shuang-Hui,GUO Hui-Chen,SHANG You-Jun,BAI Xing-Wen,LIU Xiang-Tao and XIE Qing-Ge.Eukaryonization of T7 RNA Polymerase Prokaryotic Expression System and Development of Its Couple Expression System[J].Chinese Journal of Biotechnology,2007,23(5):947-952. |
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| Authors: | ZHENG Hai-Xue JIN Ye YIN Shuang-Hui GUO Hui-Chen SHANG You-Jun BAI Xing-Wen LIU Xiang-Tao and XIE Qing-Ge |
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| Institution: | State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;Graduate School,;State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China;State Key Laboratory of Veterinary Etiological Biology, Key Laboratory of Animal Virology of Ministry of Agriculture, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Lanzhou 730046, China |
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| Abstract: | |
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| Keywords: | IERS |
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